Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, 100193, China.
Institute of Bioinformatics and Medical Engineering, School of Electrical and Information Engineering, Jiangsu University of Technology, Changzhou, 213001, China.
Sci Rep. 2016 Sep 16;6:33477. doi: 10.1038/srep33477.
HIV-1 integrase (IN) plays an essential role in viral replication and thus serves as an important target for chemotherapeutic intervention against HIV-1 infection. However, the current three clinical IN inhibitors, raltegravir, elvitegravir and dolutegravir share the same inhibitory mechanism, resulting in a common clinical resistance profile which have emerged in infected patients receiving treatment. Therefore, it is important to develop small molecule inhibitors that impair IN function with distinct mechanisms of action. In this work, a magnetic-beads based biochemical assay targeting the protein-protein interaction (PPI) between HIV IN and the cellular cofactor LEDGF/p75 was developed for identification of HIV-1 IN inhibitors. Furthermore, a library containing 1000 US. Food and Drug Administration (FDA)-approved drugs currently used for human medication was screened to identify inhibitors targeting the PPI. The assay was proved to be quite robust and with the novel assay we successfully identified dexlansoprazole (IC50 of 4.8 μM), a FDA-approved proton pump inhibitor, as a potential inhibitor for the PPI between IN and LEDGF/p75, which bound to the LEDGF/p75 partner with a kinetic dissociation (Kd) constant of 330 nM ± 2.6 nM.
HIV-1 整合酶（IN）在病毒复制中起着至关重要的作用，因此是抗 HIV-1 感染化学治疗干预的重要靶标。然而，目前三种临床 IN 抑制剂（raltegravir、elvitegravir 和 dolutegravir）具有相同的抑制机制，导致接受治疗的感染患者出现共同的临床耐药谱。因此，开发具有不同作用机制的小分子抑制剂来损害 IN 功能非常重要。在这项工作中，开发了一种基于磁珠的生化测定法，针对 HIV IN 与细胞共因子 LEDGF/p75 之间的蛋白-蛋白相互作用（PPI），用于鉴定 HIV-1 IN 抑制剂。此外，筛选了包含 1000 种美国食品和药物管理局（FDA）批准的用于人类用药的药物的文库，以鉴定针对 PPI 的抑制剂。该测定法被证明非常稳健，并且我们使用新型测定法成功地鉴定出 dexlansoprazole（IC50 为 4.8 μM），一种 FDA 批准的质子泵抑制剂，作为 IN 和 LEDGF/p75 之间 PPI 的潜在抑制剂，与 LEDGF/p75 伙伴的结合动力学解离（Kd）常数为 330 nM ± 2.6 nM。
